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Four recently synthesized cationic zinc(II)-benzonaphthaporphyrazines 1–4 were studied in vitro and in vivo for their photodynamic therapy (PDT) effectivity. The photophysical measurements showed that in solution and in Cremophor micelles all examined compounds exhibit very high absorption intensity in the spectral range between 680 to 750 nm. The fluorescence emission for 3 and 4 was very well expressed in different media, as well as in cell culture. The dark toxicity examinations on invasive human bladder carcinoma cell line EJ did not show any traces of toxicity. The investigations connected with the detection of their phototoxic capacity on the same cell line demonstrated very promising results especially with photosensitizers 3 and 4. The in vivo studies with these two compounds demonstrating high cell-phototoxic effect were carried out against Lewis lung carcinoma in mice after incorporation in Cremophor micelles. The excitation was done at the respective maximum absorption wavelength for each of the sensitizers at a fluence rate of 380 mW cm^-2 and a fluence of 360 J cm^-2. The phototherapeutic effect was evaluated through macroscopic observations (tumour growth delay) and by electron microscopy detection. According to these approaches the best effect (including tumour destroyment) was detected after PDT treatment with the cationic tribenzonaphthoporphyrazinato-zinc(II) 3. Typical features of random, but not of programmed, cell death necrosis were observed.

The antimicrobial activity of new meso-tetrakis(N-methyl-6-quinolinyl)-substituted porphyrins and meso-tetrakis(N-methyl-6-quinolinyl)-substituted chlorins is described. The dark toxicity and photosensitising potentials of free-base (TQP and TQC) and its Sn(IV)-complexes [(TQP)Sn(IV) and (TQC)Sn(IV)] were tested on Gram-positive (Staphylococus aureus), Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria and two species of yeasts (Candida albicans and Rhodotorula bogoriensis). The results described in this paper show that TQP and (TQP)Sn(IV) did not inhibit the growth of S. aureus in the dark, but efficiently photosensitize the inactivation of this Gram-positive bacteria. These porphyrins have no appreciable photosensitizing activity towards Gram-negative bacteria. However, (TQP)Sn(IV) shows high dark toxicity against E. coli and P. aeruginosa. The free-base derivatives demonstrated dark activity only in the case of P. aeruginosa. We suppose that these meso-tetrakis(N-methyl-6-quinolinyl)-substituted porphyrins can bind to the Gram-negative bacteria outer membrane receptors that transported vitamin B₁₂. The meso-substituted chlorins TQC and (TQC)Sn(IV) have shown similar efficiency in the dark- and photoinactivation of S. aureus. They revealed a middle level of dark toxicity towards Gram-negative bacteria. The Sn(IV)-complex of chlorin in comparison with free base and metalloporphyrins are more effective in photoinactivation of Gram-negative bacteria. Yeasts, such as Candida albicans and Rhodotorula bogoriensis are more sensitive to photodynamic inactivation as bacterial cells. The effects of (TQP)Sn(IV) and (TQC)Sn(IV) are more expressed than effects of free bases.

The effect of three amino acid protoporphyrin IX (PPIX) derivatives evaluated as sensitizers in photodynamic therapy — disodium N,N-dialanyl protoporphyrinate (PP(Ala)₂Na₂), diarginine protoporphyrinate (PPArg₂) and diarginine N,N-dialanyl protoporphyrinate (PP(Ala)₂Arg₂) – non-irradiated and pre-irradiated with UV-A, on respiratory burst of non-stimulated and opsonized zymosan stimulated neutrophils was studied. A potential synergistic effect of diazepam (7-chloro-1-methyl-5-phenyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one) was also examined. PP(Ala)₂Na₂ showed strong pro-oxidant effect towards non-stimulated neutrophils, while PPArg₂ and PP(Ala)₂Arg₂ revealed no significant effect. In the case of stimulated neutrophils all studied porphyrins showed antioxidant effect, although for PP(Ala)₂Na₂ this effect was significantly weaker than that of PPArg₂ and PP(Ala)₂Arg₂. After pre-irradiation with UV-A (λ = 365 nm, fluence 2.0 J.cm^-2, fluence rate 6.7 mW.cm^-2) the antioxidant activity of all studied sensitizers towards non-stimulated granulocytes did not change significantly when compared to effects of non-irradiated porphyrins, while in the case of stimulated cells, only PPArg₂ caused significant decrease of respiratory burst. Non-irradiated diazepam showed significant antioxidant effect and enhanced antioxidant effect of all studied porphyrins towards stimulated neutrophils, while after UV-A pre-irradiation it revealed no significant antioxidant effect on non-stimulated and stimulated neutrophils, both alone and in combination with porphyrin sensitizers.

Synthesis of pyropheophorbide-a-paclitaxel (PPa-PTX) conjugate was performed in high yield with the aim of searching for an optimal agent for cancer treatment. After synthesis, the conjugate was confirmed to be linked through an ester bond at the 2′ position of the paclitaxel moiety using multi-nuclear magnetic resonance spectroscopy. Phototoxicity of PPa and PPa-PTX conjugate, as well as PTX, was evaluated with three human cancer cell lines (HeLa, CaSki and TC-1). The new conjugate at 0.01–0.06 μM displayed 20–40% higher phototoxicity in HeLa and CaSki cell lines than free PPa and PTX. Furthermore, cellular uptake of these bio-molecules was examined by confocal laser scanning microscopy. Although PPa-PTX showed a delayed uptake compared to PPa, it penetrated completely into cells within 24 h incubation.

In order to improve the therapeutic effect of zinc phthalocyanines (ZnPc), a photoactive nanodrug was prepared with acetylated chondroitin sulfate (AcCS), utilizing a simple chemical method. AcCS/ZnPc nanodrugs have a unimodal size distribution below 200 nm and a negative surface charge due to AcCS located on the nanodrug surface. In organic solvent such as DMSO or DMF, it has strong fluorescence intensity and generates abundant singlet oxygen. However, in aqueous solvent, AcCS/ZnPc nanodrugs developed a self-organized form which induced reducing fluorescence intensity and singlet oxygen generation. The cellular uptake of the nanodrug was determined using a cell lysis test and confocal microscopy observation. The results indicated that cellular internalization efficiency of the nanodrug was 1.7–2.1 times higher than that of free ZnPc. Also, the phototoxicity of the nanodrug was detected via MTT assay with or without light. Although free ZnPc did not exhibit cytotoxicity in both light and dark condition, the nanodrug exhibited increasing cytotoxicity after irradiation. We therefore suggest that AcCS/ZnPc nanodrugs may have promising applications as new photodynamic agents for the clinical treatment of various tumors.

Photodynamic therapy (PDT) is approved for clinical indications including several (pre-) cancers of the skin and solid tumors of the brain and the gastrointestinal tract. It operates by an acute cellular response caused by oxidation of cell components following light-induced and photosensitizer-mediated generation of reactive oxygen species. By this, PDT is capable of inducing the major types of cytotoxic responses: autophagy, apoptosis, and necrosis. As excited photosensitizer molecules react rather non-specifically with neighboring molecules, we suggest that with PDT and most (if not any) cell-localizing photosensitizers, all kinds of cellular responses can be provoked — following a strict dose-dependency, i.e. a transition from survival, over apoptosis to necrosis depending on the applied photosensitizer concentration or light dose. In this review, we briefly discuss (i) the types of cell death induced by PDT focusing on apoptosis induction, (ii) a simple experimental approach to quickly assess the dose-dependent phototoxic responses based on viability assays, and (iii) an overview of in vitro apoptosis detection methods for further in depth analyses. With this conceptual framework, we attempt to provide a rational experimental approach for initial in vitro, cell-based characterization of newly synthesized photosensitizers or formulations thereof — thus to plug the gap between subsequent in vivo evaluation and the preceding fundamental (physico-)chemical work devoted to the improvement of photosensitizing drugs based on mainly porphyrins, phthalocyanines and their derivatives.

Three imidazolyl-appended porphyrins were synthesized and characterized by ¹H NMR, elemental analyses, MS and UV-vis spectra. Anticancer activities of porphyrins have been evaluated against cutaneous squamous cell carcinoma (A431 cells) in vitro. The results indicate that the porphyrins have high selective cytotoxicity towards A431 cells in the absence of light and improved phototoxic activity upon exposure to UV light. The yield of singlet oxygen generated by porphyrins were also evaluated by measuring the absorption decay of 1,3-diphenylisobenzofuran (DPBF) in DMF. The phototoxicities of porphyrins against A431 cells were enhanced along with the increase of singlet oxygen.

One novel porphyrin 5,10,15-tris(phenyl)-20-[4-(2-(2-methyl-5-nitro-imidazolyl)ethoxyl)phenyl] porphyrin and its zinc(II) metalloporphyrin were synthesized and characterized by IR, UV-vis, ¹H NMR, MS and elemental analysis. The single crystal structure of zinc(II) porphyrin shows that the Zn(II) ion is coordinated with four nitrogen atoms of porphyrin ring and one oxygen atom of ethanol from axial, forming a five-coordinated square pyramidal geometry. Their cytotoxicity and photodynamic activity against breast cancer cells were studied. The results indicate that both of the porphyrins display high phototoxicity to the breast cancer cells with the negligible dark toxicity. In addition, the photodynamic activity of zinc(II) porphyrin was obviously higher than that of the free porphyrin.

Photodynamic inactivation of microorganisms known as antibacterial photodynamic therapy (APDT) is one of the most promising and innovative approaches for the destruction of pathogenic microorganisms. Among the photosensitizers (PSs), compounds based on cationic porphyrins/metalloporphyrins are most successfully used to inactivate microorganisms. Series of meso-substituted cationic pyridylporphyrins and metalloporphyrins with various peripheral groups in the third and fourth positions of the pyrrole ring have been synthesized in Armenia. The aim of this work was to determine and test the most effective cationic porphyrins and metalloporphyrins with high photoactivity against Gram negative and Gram positive microorganisms. It was shown that the synthesized cationic pyridylporphyrins/metalloporphyrins exhibit a high degree of phototoxicity towards both types of bacteria, including the methicillin-resistant S. aureus strain. Zinc complexes of porphyrins are more phototoxic than metal-free porphyrin analogs. The effectiveness of these Zn–metalloporphyrins on bacteria is consistent with the level of singlet oxygen generation. It was found that the high antibacterial activity of the studied cationic porphyrins/metalloporphyrins depends on four factors: the presence in the porphyrin macrocycle of a positive charge (+4), a central metal atom (Zn${}^{2+})$ and hydrophobic peripheral functional groups as well as high values of quantum yields of singlet oxygen. The results indicate that meso-substituted cationic pyridylporphyrins/metalloporphyrins can find wider application in photoinactivation of bacteria than anionic or neutral PSs usually used in APDT.

Photodynamic inactivation of microorganisms known as antibacterial photodynamic therapy (APDT) is one of the most promising and innovative approaches for the destruction of pathogenic microorganisms. Among the photosensitizers (PSs), compounds based on cationic porphyrins/metalloporphyrins are most successfully used to inactivate microorganisms. Series of meso-substituted cationic pyridylporphyrins and metalloporphyrins with various peripheral groups in the third and fourth positions of the pyrrole ring have been synthesized in Armenia. The aim of this work was to determine and test the most effective cationic porphyrins and metalloporphyrins with high photoactivity against Gram negative and Gram positive microorganisms. It was shown that the synthesized cationic pyridylporphyrins/metalloporphyrins exhibit a high degree of phototoxicity towards both types of bacteria, including the methicillin-resistant S. aureus strain. Zinc complexes of porphyrins are more phototoxic than metal-free porphyrin analogs. The effectiveness of these Zn–metalloporphyrins on bacteria is consistent with the level of singlet oxygen generation. It was found that the high antibacterial activity of the studied cationic porphyrins/metalloporphyrins depends on four factors: the presence in the porphyrin macrocycle of a positive charge (+4), a central metal atom (Zn²⁺) and hydrophobic peripheral functional groups as well as high values of quantum yields of singlet oxygen. The results indicate that meso-substituted cationic pyridylporphyrins/metalloporphyrins can find wider application in photoinactivation of bacteria than anionic or neutral PSs usually used in APDT.